High Performance Liquid Chromatography Using UV Detection for the Simultaneous Quantification of the New Non-nucleoside Reverse Transcriptase Inhibitor Etravirine (TMC-125), and 4 Protease Inhibitors in Human Plasma

Abstract

Etravirine (TMC-125, ETV) is a second-generation nonnucleoside reverse transcriptase inhibitor (NNRTI) that demonstrates potent activity against NNRTI-resistant strains of human immunodeficiency virus type-1 (HIV-1). According to the DUET studies (randomized, double-blind, placebocontrolled trials), overall, ETV was well tolerated in treatment-experienced patients infected with HIV-1, with its safety and tolerability profile generally comparable to placebo at week 24.1,2) Additionally, 48-week data pooled from these studies showed greater virologic and immunologic responses compared with placebo. 3)In the latest HIV treatment, ETV has been used in combination with ritonavir-boosted protease inhibitor (PI) and integrase inhibitor for therapy-experienced HIV-1-infected patients. On the other hand, as ETV is a substrate and inducer of cytochrome P450 3A4 (CYP3A4), ETV may induce metabolism of PI and alter the concentrations of co-administered PIs. 4) In order to ensure optimal drug efficacy and prevention of resistance, it is essential to monitor plasma concentrations of ETV and PIs.Fayet et al. 5) and Quaranta et al. 6) succeeded in determining plasma concentrations of ETV and other drugs through the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS). Rezk et al. 7) have also developed a method to measure plasma concentrations of ETV and PIs by LC-MS. LC-MS or LC-MS/MS assay is very sensitive and accurate. However, MS equipment is very expensive and unavailable in conventional hospital laboratories. Therefore, development of alternate methods is necessary.Recently, D'Avolio et al. 8) reported a new HPLC method that employs a photo diode array (HPLC-PDA) for quantification of ETV and other antiretroviral drugs. This method is simple, reliable, and sensitive, using cost-effective instrumentation when compared with others. 5-7) However, this method requires a solid phase extraction. Furthermore, in general hospitals, a UV detector coupled with HPLC is more popular than a PDA detector. The HPLC-UV method is a user-friendly assay that is readily adaptable to standard laboratory equipment for routine therapeutic drug monitoring (TDM).In this study, we propose the simultaneous quantitative assay of ETV and 4PIs, darunavir (DRV), atazanavir (ATV), ritonavir (RTV) and lopinavir (LPV) in a simple procedure that is derived from a previously established HPLC-UV method.9) This method can be applied to pharmacokinetic studies of PIs and ETV, the newest NNRTI, and it is useful when evaluating the clinical significance of TDM for these drugs. Etravirine (TMC-125, ETV) is a second-generation non-nucleoside reverse transcriptase inhibitor (NNRTI) that demonstrates potent activity against NNRTI-resistant strains of human immunodeficiency virus type-1 (HIV-1). Thus, ETV has been used in combination with ritonavir-boosted protease inhibitor (PI) and integrase inhibitor for therapy-experienced HIV-1-infected patients. On the other hand, as ETV is a substrate and inducer of cytochrome P450 3A4 (CYP3A4), ETV may induce ...

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